The Ohio State University
Nelly Arguello-Blanco, Mao Huang
Genomic selection facilitates rapid cycling through a breeding cycle by eliminating the need to phenotype prior to selecting superior parents and crossing among them. In winter wheat we can now complete a cycle of GS in about 12 months and two greenhouse seasons. Season consists of planting F1s from the previous cycle and selfing to obtain F2 seed. The second season involves planting and genotyping the F2s, predicting their value with GS, selecting and crossing the best, and harvesting the F1 seed. Our breeding program has completed five cycles of GS in one population primarily for grain yield, over the past five years. We have completed three cycles of GS for resistance to Fusarium Head Blight in a second population. Genotyping was done using genotyping-by-sequencing. This provides an opportunity to assess the changes in the population that have occurred as a result of this rapid cycling. These include 1) changes in genomic estimated breeding values for grain yield and FHB resistance, 2) effect of selection and drift on allele frequencies including fixation, 3) effect of selection on diversity and genetic relationships, and 4) changes in linkage disequilibrium. We are conducting these analyses and will present the results.
Crop Diseases Research Institute, PARC Substation, Murree Pakistan
Sufyan,Muhammad, Abid Majeed, Satti, Munir, Anjum, Fayyaz, Muhammad, Atiq ur Rehman, Rattu, Imtiaz, Muhammad, , , , , , , , , , , , , , , , , ,
225 Puccinia striiformis f.sp. tritici isolates collected from wheat growing areas of Pakistan during 2013-2016 were analyzed using 18 near isogenic yellow rust differentials. Seventy eight races were identified among collection in which 20 were common (n > 2). Rest of the races were very rare and encountered only once (n=1). Races 574212, 574232, 474232, 474233, 574213 and 434232 were most frequent (n> 15). Pathogenic diversity analysis of the collection reveal high diversity (H =3.57) of the P. striiformis population of pakistan. On the basis of phenotypic response to yellow rust genes, the most frequent races could be grouped into 5 diverse groups. Distinct grouping was also observed in rarely encountered isolates. Most of the races were highly complex and 80% isolates had complexity ranging from 8 to 11. Virulence frequency for Yr6, Yr7, Yr8, Yr17, Yr27, Yr43 & YrExp2 remained above 80% while that of Yr1, Yr9 and Yr44 remained over 40%. Partial virulence was detected for Yr5, while virulence to Yr10, Yr15, YrSP was found in < 4% isolates. Paper discuss spatial and temporal distribution of P. striiformis races in Pakistan.
Ravi Singh, Karim Ammar
Stripe rust, caused by Puccinia striiformis tritici (Pst), continues its evolution towards virulence to race-specific resistance genes. Identification of Mexican Pst isolates MEX16-03 and MEX16.04 that changed infection types of Yr10 testers from 1 to 9 and for Yr24 (=Yr26) testers from 3 to 9 indicated that a mutation for virulence to these resistance genes has occurred in a predominant race detected in 2014 and maintained at CIMMYT as MEX14.191 and at INIFAP as CMEX14.25. Isolate MEX14.191 was responsible for the susceptibility of popular varieties Nana F2007 and Luminaria F2014 grown in central Mexican highlands. Isolate MEX16.04 has the following avirulence/virulence formula: Yr1, 5, 15, SP/Yr2, 3, 6, 7, 8, 9, 10, (17), 24, 26, 27, 28, 31, 32 using the Avocet near-isolines and other known testers. Virulence to Yr10 and Yr24 (=Yr26) were also confirmed by testing seedlings of cultivars Moro (Yr10), Chuanmai 42, and Neimai 836 (Yr24). Seedling tests carried on 200 bread wheat, 550 durum, and 460 synthetic hexaploid wheats with their respective durum parents from CIMMYT collection indicated that MEX16.03 and MEX16.04 do not represent a major threat because a majority of the lines remained resistant to these isolates. However, it is worth mentioning that durum cultivars, such as Khofa, Desert King, Anatoly, Movas, and Llareta INIA, and 10 primary synthetic hexaploid or synthetic-derived bread wheats that were resistant to MEX14.191 became susceptible to MEX16.03 and MEX16.04. Our results indicate that resistance gene Yr10 was absent and Yr24 occurred in low frequency in CIMMYT bread wheat germplasm. A majority of CIMMYT durum wheat possibly carried Yr24 in combination with other effective gene(s).
Scuola Superiore Sant'Anna
Yosef G.,Kidane, Cherinet, Alem, Bogale, Nigir, Dejene, Mengistu, Carlo, Fadda, , , , , , , , , , , , , , , , , , , ,
The Ethiopian plateau hosts thousands of durum wheat landraces cultivated in low input agriculture conducted by an estimated 70 million smallholder farmers. Having thoroughly characterized the phenotypic and molecular uniqueness of Ethiopian durum wheat landraces, we produced a large nested association mapping (NAM) population harnessing their mostly untapped diversity in a set of recombinant inbred lines (RIL). The NAM founders are 50 landraces providing valuable traits such as drought tolerance and resistance to pests, and maximizing molecular diversity. Each selected landrace was crossed to a durum wheat line with an international background (Asassa), establishing independent interconnected bi-parental families, for a total of 6,280 RILs currently in F8. The Ethiopian NAM is at once i) a powerful QTL mapping tool that will side the increasing availability of genomic tools in wheat towards high-throughput candidate genes identification, and ii) a large pre-breeding panel closing the gap between local and international materials. Here we discuss the molecular and phenotypic characterization of twelve NAM families, represented by 100 RILs each. The 1,200 NAM RIL showed elevated allelic variation and a genetic structure reminiscent of the breeding design followed. The NAM RILs were phenotyped for ten agronomic and five disease traits in multiple locations in the Ethiopian highlands. A quantitative method eliciting smallholder farmers traditional knowledge was used to record local farmers appreciation of NAM RILs in all phenotyping locations. We report that the superior genetic properties of the NAM can be used to map QTL for both agronomic and farmer traits with unprecedented precision. The most promising NAM RILs can be identified combining farmers appreciation and agronomic measures, and prioritized for introgression of Ethiopian landraces traits in breeding pipelines aiming at higher uptake and productivity in local agriculture.
ICAR NRC Plant Biotechnology
The present challenge in wheat breeding is to decipher the molecular mechanisms of heat stress response and thermotolerance in detail for future applications. Several reports indicate the ability of plants to maintain a memory of stress exposure throughout their ontogenesis and even transmit it faithfully to the following generation. Here, three diverse genotypes of wheat viz., HD2967, WR544 and C306 were used for thermotolerance assays. The genotype HD2967 was able to withstand heat stress regimes (37?C and 42?C, 2 hours). Harvested seeds were sown and further raised for two consecutive years and phonotypical data evaluated in natural field condition by exposing to heat stress during generative stages in a heat trap chamber. Maximum tiller numbers and flag leaf length were noticed in second generation plant of 37?C heat exposure whereas flag leaf width in second generation 42?C heat exposure relative to the untreated plant. Auricle length showed no difference but plant height was notably increased in the second year in all the heat exposed plants. In grain yield index, ear head length was greater in the second year and fluctuations in grain number was noticed among the heat treated plant with more yields in 42?C and 42?C HTHT in the second year. IRGA and SPAD recording showed high photosynthesis and chlorophyll content in 37?C HTHT. High modulation of transcripts of several genes involved in DNA methylation and heat stress were also observed. The domino effect of heat stress in earlier generation, in this transgenerational analysis, points towards a probable epigenetic effect. Further studies are in progress to confirm and clarify the mechanisms for future manipulation in breeding for thermotolerance.
International Maize and Wheat Improvement Center (CIMMYT), P.K. 39 Emek 06511 Ankara, Turkey
Nilufer,Akci, Sridhar, Bhavani, Mesut, Keser, Fatih, Ozdemir, Ruth, Wanyera, Alexey, Morgounov, , , , , , , , , , , , , , , , , ,
A diverse set of winter wheat germplasm was screened for resistance to stem rust in large-scale trials in Kenya and Turkey during 2009-16. The study aimed to select resistant material and characterize types of resistance and possible genes, as well as evaluate agronomic traits and resistance to other diseases to select superior variety candidates and parental lines. The study material was comprised of various Facultative and Winter Wheat Observation Nurseries (FAWWON), which are developed and distributed by the International Winter Wheat Improvement Program (www.iwwip.org) in Turkey. More than 1600 global accessions were screened, with most evaluated for two years. Based on stem rust data from Kenya, more than 400 genotypes were identified exhibiting adequate levels of resistance to the Ug99 race group. The highest number of resistant lines originated from IWWIP (~170), USA (~100), Russia (~40), Iran (~30), Romania (~20), and South Africa (~20). Material was also tested at two sites in Turkey: Haymana (artificial inoculation) and Kastamonu (natural infection). There was no significant correlation between stem rust severities in Kenya and in Turkey, due to differences in stem rust pathotypes. However, a set of germplasm (more than 100 entries) has been identified as resistant in both countries. This set represents promising material as variety candidates and parental lines; another study is currently identifying the genes controlling the stem rust resistance in this population. IWWIP distributed stem rust resistant germplasm to its global collaborators during 2010-2015, in response to the threat from the Ug99 race group. New resistant germplasm combining broad adaptation, high yields, and resistance to other diseases is available on request.
Seed and Plant Improvement Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran
The basidiomycetous fungus, Puccinia graminis f. sp. tritici (Pgt) causes stem rust disease as one of the most destructive wheat pathogens, worldwide. TTKSK and other Pgt races under Ug99 race group are considered as major threats to wheat production in east Africa and CWANA region by defeating the stem rust resistance gene Sr31, while its ineffectiveness was reported in Iran in 2007. Race TKTTF of Pgt caused a severe stem rust epidemic in southern Ethiopia in 2013, and was spread to Europe through 2016 Sicily outbreak. This research describes race identification of Iranian isolates collected during the widespread distribution of stem rust in 2014-16. Purified urediniospores of 123 Pgt isolates were separately inoculated on seedlings of 20 North American differential wheat cultivars carrying different Sr resistance gene/s. Infection types were recorded at 14 days post inoculation (dpi) using Stakman et al. 0-4 scale. Based on the letter code nomenclature, we identified the Pgt races TKTTF, TTTTF, TTKSK, TTKTK, PKTTF, TKSTF, PKSTF, PKTTC, PTRTF, PTTTF, PKSTC, TTRTF, TKSTC and PKRTF in Iran. TKTTF and TTTTF were determined as prevalent Iranian Pgt races. This is the first report of race TTKTK, a new variant of Ug99 race group with virulence on Sr31 and SrTmp resistance genes, in Iran. Since TTKTK primarily occurred in south west of Iran, the migration route for this new race seems to be similar to race TTKSK. The high race variation observed in this study could indicate a high genetic diversity among P. graminis f. sp. tritici populations in Iran, as a wheat center of origin.
Department of Botany, Faculty of Biology, Government College Murree, Pakistan
Barkat Ali, Muhammad Fayyaz, Atiq ur Reman Rattu, Abdul Samad Mumtaz, Muhammad Imtiaz
The new arrival of wheat rust pathotypes through migration during wheat cropping season requires regular monitoring to secure wheat production. In the present study, we collected leaf rust (Puccinia triticina Eriks.) infected wheat leaves from three major wheat growing provinces of Pakistan in the year 2014 to assess the haplotype diversity of P. triticina (Pt) isolates. The rDNA ITS sequence data of collected isolates was used in NCBI BLAST analysis. The blast hits showed best matches with Pt accessions EU014050 (Iran), JN120331 (Iran), JX533577 (Iran), AY956549 (Iran), DQ417412 (Czech Republic), DQ417418 (Israel), DQ417413 (Slovakia) and AF511083 (Louisiana). However, in cluster analysis, the Pakistani isolates showed strong bootstrap support with only Iranian and Indian (races 77-5 & 104-4) accessions that indicated eastward migratory mode of Pt pathotypes in Pakistan through westerly wind patterns. The predominant genotype DQ417412 (similar in alignment with AY956549 from Iran) overcome the resistance of top Pakistan mega varieties Seher06, Inqilab91, Kiran95, SKD1, TJ83 and NIFA-Batoor. Hence, the ITS based information remains a rapid molecular tool for pathogen surveillance across countries and continents.
ICAR Indian Institute of Wheat and Barley Research, Karnal
Satish Kumar, Rekha Malik, Garima Singhroha, Vinod Tiwari, Gyanendra Pratap Singh
Breeding rust resistant cultivars using conventional methods is time-consuming, complex and slow, but molecular markers offer a rapid alternative for developing cultivars with improved disease resistance. Three wheat cultivars, DBW88, DBW107, and DBW110, from different production zones were used as recipients for incorporation of resistance genes using a marker-assisted backcross (MAB) breeding approach. Leaf rust resistance gene Lr32 is being incorporated into all the three varieties, stripe rust resistance gene Yr15 is being incorporated into DBW88 and DBW107, and stem rust resistance gene Sr26 is being added to variety DBW110. Lines PBW703 (Yr15), FLW15 (Lr32) and Avocet (Sr26) were used as donors. Six cross combinations viz., DBW88/PBW703, DBW107/PBW703, DBW88/FLW15, DBW107/FLW15, DBW110/FLW15 and DBW110/Sr26 were made at Karnal during 2015-16 and the crosses were grown at IIWBR-RS, Dalang Maidan for backcrossing. BC1F1 plants were raised at Karnal during 2016-17. Both foreground and background selections were practiced in each combination. SSR markers gwm264 and barc135 were used for foreground selection of Lr32, marker barc8 was used for selection of Yr15, and markers Sr26#43 and BE518379 were used to detect presence and absence of Sr26. From 90 to 127 polymorphic SSR markers chosen for each cross from an initial set of 800 screened on the parents are being used for background selection.
Ethiopian Institute of Agricultural Research
Bedada Girma, Endale Hailu, Fikrte Yirga, Bekele Abeyo, Gordon Cisar, Gina Brown-Guedira, Erena Edae, Pablo Olivera, Matthew Rouse
In Ethiopia, breeding resistant wheat varieties is a priority for wheat rust management although new virulent rust races have periodically resulted in losses of R-genes, epidemic outbreaks, and yield losses of up to 100%. During 2014 and 2015, 160 wheat varieties and lines including five checks and 12 differential lines with known resistance genes were evaluated against four stem rust pathogen races at both seedling and adult plant stages. In the field at Kulumsa, Ethiopia, the lines were evaluated in four separate nurseries in an augmented design where each nursery was inoculated with a different Puccinia graminis f. sp. tritici race: TTKSK, TKTTF, TRTTF, and JRCQC. Kingbird, a check variety, displayed low average Area Under the Disease Progress Curve (AUDPC) (67 to 238) and Average Coefficient of Infection (ACI) (1.1 to 9.7) in response to the four races. Effect of lines possessing Sr24+Sr36 and Sr31+Sr36 resistance genes on rust development was comparable to Kingbird or even better. Likewise, 48, 34, 19 and 28 varieties and lines had lower or comparable AUDPC for TTKSK, TKTTF, TRTTF and JRCQC compared to Kingbird. Commercial bread wheat varieties Shorima, Huluka, Hogana, and advanced lines CIMMYT 14, ETBW7058, ETBW7101 and ETBW7258 for which Sr24/Lr24 was postulated, possessed AUDPC and ACI lower than Kingbird. However, all these lines possessed susceptible or intermediate seedling reactions to Sr24-virulent race TTKTT from Kenya at the seedling stage. CIMMYT 18 showed susceptibility to 3 races at seedling stage, but lower AUDPC and ACI than the checks except Kingbird, indicating adult plant resistance. However, this adult plant resistance was marginal in effect to race TKTTF. Resistance genes Sr2, Sr57/Lr34, Sr24/Lr24, Sr25/Lr19, Sr38/Lr37 and SrTmp were postulated at various frequencies in this germplasm. Seedling and adult plant resistance sources identified can be used for rust resistance breeding in Ethiopia.